LOCUS       sequence-385261-        6896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc-Eps8L2-FL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6896)
  AUTHORS   Calero-Cuenca FJ, Osorio DS, Carvalho-Marques S, Sridhara SC, 
            Oliveira LM, Jiao Y, Diaz J, Janota CS, Cadot B, Gomes ER
  TITLE     Ctdnep1 and Eps8L2 regulate dorsal actin cables for nuclear 
            positioning during cell migration.
  JOURNAL   Curr Biol. 2021 Apr 12;31(7):1521-1530.e8. doi: 
            10.1016/j.cub.2021.01.007. Epub 2021 Feb 9.
  PUBMED    33567288
REFERENCE   2  (bases 1 to 6896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cub.2021.01.007"; journalName: "Curr Biol"; date: 
            "2021-04-12- 12"; volume: "31"; issue: "7"; pages: "1521-1530.e8"
COMMENT     Sequence Label: pRK5mycGW-Eps8L2
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        158..349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     306..326
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    418..552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(442..461)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     496..515
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    593..617
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2762..2786)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2805..2834)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2883..2902)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2915..2933)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2916..2933)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3143..3167)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3164..3367)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3193..3213)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3368..3747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3776..3792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3776..3792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3789..3811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3800..3816
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3824..3830,3831..3848,3849..3854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3869..3890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4007..4024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4178..4766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4258..4277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4937..5728,5729..5797))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5560..5579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5798..5902)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5970..5988
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6026..6048)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6148..6167
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6184..6639)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6321..6342)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6533..6552
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6837..6859
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6851..6868
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6852..6868
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(6888..6896,1..349))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
       61 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      121 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      181 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      241 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      301 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
      361 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgccgaatt aattcccgat
      421 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
      481 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
      541 aataaacaag ttgggccatg gcggccaagc ttctaatcta gaattcccat caaccacttt
      601 gtacaagaaa gctgggtgct gtcctccccc ctcctctgat tcatggaatg aaacttgttc
      661 atgagttctt ccagctccga cccactttgc tgcttctcca ggaaggcctt ctgcatggtg
      721 agctggctgt acacgcggac gccctcctcg ccgcacactt tcttcagctc ctccttgttg
      781 agggagaaga gctgcggccc ggtcaggatg cccaggttct ccacgatccg cgggctgaag
      841 gccttggctt ccagccaggc gcggacctcg tccggacccg actcgtaggt gagcggctgg
      901 ctcacgggct ggctgcgctc cacgcggaag tgcctctgtg gctgcgccct tatattcgat
      961 atcttgcgga tgagctcgtc gttgacctcg tccatgtgct gcatgagttc gtccttattg
     1021 cccgggaagc ttgggggtag cttgtgggtc gggctggcgg ggccccagta cttctgaccg
     1081 gcctgctcga acggggcgcc ggcgtcctcc ggtcgcgcct cgcctaggat gttgcagggc
     1141 acgtaccccg cctggccgct gcggctgcgc agcttccacc actgccggcc gtcctccagc
     1201 acctctagga cctcatcctt gagcaccgat agctcgttgg catttcgggc tgtgaagtca
     1261 tagagaattt tcacatattt ggccatggct ggtgttggct ggtagcccct gtgagtatgt
     1321 ggggagctga ctggtggtag ggcatccccc gggggggtgg gctctgaagt ggggctgtgc
     1381 ttctgggagt tccggatact ttgcctcgac actggactca cctcctcgat gggggcagac
     1441 gccagcccct ccacctccca gggggcctcc tgcagcacat ccacaggagg ctcccagccg
     1501 ctgtggaact tgggcacgta gaggggcacc tgtggctccc gcggccactc ggaacggggc
     1561 cgcatccagc tctctcccag tgactcccac agcgacatct ccttagggac caggtggccg
     1621 cgcaggaagt ccacggcatc tcgggagagc agtgggcagg agacggagcg tgcgatgtct
     1681 gggccactgc aggtgttgac gatcaggtcc agaggcccga agaggaagtg cacgagctcc
     1741 gcggcgctgg ggttctggat gtgcttctgc agctttgcca gcaagttaat cgccagcttg
     1801 attttctgga agcagtcgat gaactcgccc tcagaggggg gccgtgcccg cagtgtgagg
     1861 acgccctctg ctggcgcctt cttgcccttc ttcttcccct ttttccgctg gttcagctgc
     1921 ttgaaagcct cggctgcctt ctgcagccgg gccacaaacc actcgatgtc gtccagggcg
     1981 cagttgagga tttgcgtctc cttctctatc ttctgagcca gcacggcccg cggctcctcc
     2041 tgcgactccc gacggcggaa acctggctcg ctgagtggca cctgcgggcc cacgcgattc
     2101 ttggcctccg gggaatcccc gccgcggtgc tggaagggga tgggcgccgg gccctgggga
     2161 ggaggcagga tggactgccg ctgccgaatc ttctcctggt gtcccttcag ggtctgcggc
     2221 cgcatcttct tgcccagccg gcagtcggcc aacgcgctct cgatgtcctc gtgcaccagc
     2281 tctgcctcca cctcatcgca gtggaagaag tggacatccg gcttgctctg ctccgagtcc
     2341 tggcacacga gcagcagcac agacgggtag cgcagctggt tgaggaccgt ctggctgcgc
     2401 tgcaccgtgg gcagcgggaa gtcttccagc tcctcctgtg actcgatgtc cagcagccgc
     2461 agcgactggt cgttcacctg cagcagcatc tcctgggtcc agatcttctc cttggagctc
     2521 agctgcacca gcttccggat ggcgtcgtcc acagacgtga tggcttcgct cttgtccatg
     2581 atgaatgtgg ccaggtgctg gacgtggtac tgcgaggtct cgtgcatgat gacgttggag
     2641 ttggaatact tcttcctctg ctcaaacagg tccttggggc tcatcttggc cacaccgtcg
     2701 gaccggccca ggctgccatt ggtggcaccc gggcagcagc tcacggcccc ggactggctc
     2761 aagcctgctt ttttgtacaa acttgttgat ggggatccct cgagcaggtc ctcctcgctg
     2821 atcagcttct gctccatggt gggatcgata gaaccgaggt gcagttggac ctgggagtgg
     2881 acacctgtgg agagaaaggc aaagtggatg ttattctata gtgtcaccta aatcgtatgt
     2941 gtatgataca taaggttatg tattaattgt agccgcgttc taacgaagcc aagggggtgg
     3001 gcctatagac tctataggcg gtacttacgt cactcttggc acggggaatc cgcgttccaa
     3061 tgcaccgttc ccggccgcgg aggctggatc ggtcccggtg tcttctatgg aggtcaaaac
     3121 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg cttatataga
     3181 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3241 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3301 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3361 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3421 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3481 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3541 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3601 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3661 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3721 attaataact agtcaataat caatgtcggg cgagctcgaa ttaattcgta atcatgtcat
     3781 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3841 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3901 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3961 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4021 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4921 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4981 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5041 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5101 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5161 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5221 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5281 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5341 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5401 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5461 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5521 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5581 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5641 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5701 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5761 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5821 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5881 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5941 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     6001 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     6061 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6121 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6181 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6241 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
     6301 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6361 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6421 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6481 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6541 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6601 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
     6661 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6721 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6781 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6841 gtcacgacgt tgtaaaacga cggccagtgc caagctgtta acagcttttt gcaaaa
//
